Application of a MTT Assay for Screening Nutritional Factors in Growth Media of Primary Sponge Cell Culture

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• 作者：Xiaoying Zhang ; Gaë ; l Le Pennec ; Renate Steffen ; Wener E. G. Mü ; ller ; and Wei Zhang
• 刊名：Biotechnology Progress
• 出版年：2004
• 出版时间：February 2004
• 年：2004
• 卷：20
• 期：1
• 页码：151 - 155
• 全文大小：150K
• 年卷期：v.20,no.1(February 2004)
• ISSN：1520-6033

文摘

Marine sponges (Porifera) are producers of the largest variety of bioactive compoundsamong benthic marine organisms. In vitro culture of marine sponge cells has beenproposed for the sustainable production of these pharmacologically interestingcompounds from marine sponges but with limited success. The development of asuitable growth medium is an essential prerequisite for sponge cells grown in vitro.The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay wasadapted to screen for potential nutritional factors in formulating a growth mediumfor primary cell culture of Suberites domuncula. In 96-well plates, the effects ofnutritional factors including glutamine, pyruvate, iron citrate, silicon, RPMI 1640,and Marine Broth 2216 on the viable cell density were examined in primary cell cultureof S. domuncula 36 h after inoculation. Ferric iron (Fe³⁺) and pyruvate were found tosignificantly improve cell viability in a dose-dependent manner. Silicon and glutamineshowed limited improvements at certain concentrations. The supplement of RPMI 1640and Marine Broth 2216 did not increase cell viability. As a result, several improvedmedia able to maintain higher cell viability in a short-term culture of primary spongecells could be formulated.